Method for Increasing Performance of Offspring

ABSTRACT

Methods and compositions for increasing intestinal transport of nutrients or growth performance in the offspring of an animal are described. More specifically, a feed composition comprising an omega-3 fatty acid-containing composition for increasing intestinal transport of nutrients or growth performance in the offspring of the animal, and methods therefor, are described.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(e) to U.S.Provisional Application No. 60/980,143, filed Oct. 15, 2007, which isexpressly incorporated by reference herein.

FIELD OF THE INVENTION

The invention relates to methods for increasing intestinal transport ofnutrients in the offspring of an animal, and compositions therefor. Theinvention also relates to methods for increasing the growth performanceof the offspring of an animal, and compositions therefor.

BACKGROUND AND SUMMARY

Omega-3 and omega-6 fatty acids and their metabolites regulate numerousactivities in vivo, including inflammation, disease resistance, plateletfunction and vessel wall contractions. Moreover, supplementation ofomega-3 fatty acids and/or gamma-linolenic acid present in the diet ofanimals and humans are reported to have favorable effects on heartdisease, inflammatory and autoimmune disorders, diabetes, renal disease,cancer, and immunity as well as learning, visual acuity and neurologicalfunction.

On a cellular level long chain omega-3 fatty acids are readilyincorporated into the phospholipid fraction of cell membranes where theyinfluence membrane permeability/fluidity and transport. This representsa storage form of these fatty acids, where they remain until acted uponby phospholipase enzymes which release them for further conversion toeicosanoids.

Linoleic and alpha-linolenic acids are C18-containing fatty acids thatare parent compounds of the omega-6 and omega-3 families of fatty acids,respectively. Omega-3 and omega-6 fatty acids undergo unsaturation(i.e., adding double bonds) and sequential elongation from the carboxylend (i.e., adding 2-carbon units) with the D6-desaturase enzyme beingthe rate limiting enzyme in metabolism of these long chain fatty acids.The same enzymes are used for these families, making the familiesantagonistic to one another. Such antagonism, resulting fromrequirements for the same enzymes, extends into the further metabolismof the C20-containing members of these families into metabolites calledeicosanoids.

The polyunsaturated fatty acids, including omega-3 and omega-6 fattyacids, differ from the other fatty acids in that they cannot besynthesized in the body from saturated or monounsaturated fatty acids,but must be obtained in the diet. The omega-6 fatty acid, linoleic acid,is found in high quantities in vegetable oils such as corn, cottonseed,soybean, safflower and sunflower oil. The omega-3 fatty acid,alpha-linolenic acid, is found in high quantities in flaxseed oil,linseed oil, perilla oil and canola oil. Other important compoundsinclude arachidonic acid, found in animal fat; gamma-linolenic acid,found in evening primrose oil, borage oil, and blackcurrant oil; andeicosapentaenoic acid, docosahexaenoic acid, and docosapentaenoic acidderived from fish oils and algae. These long-chain fatty acids can beformed in the body by elongation and desaturation of the parent linoleicand alpha-linolenic acids if the parent compounds are supplied in thediet.

Applicants have discovered that supplementation of the diet of animalswith polyunsaturated fatty acids, including omega-3 fatty acids, derivedfrom algal sources or from non-algal sources having a highdocosahexaenoic acid content, results in positive effects for theoffspring of the animal when the mother is fed these compositionscontaining fatty acids. Interestingly, these compositions cause positiveeffects for the offspring including increased intestinal transport andincreased growth performance, including an increase in growth rate, areduced feed to weight gain, and an increase in the efficiency of feedutilization.

Methods and compositions for increasing intestinal transport ofnutrients in an offspring an animal are described herein. In oneembodiment, a method of increasing intestinal transport of nutrients inan offspring of an animal is provided. The method comprises the steps ofadministering to the animal a feed composition comprising an algalcomposition comprising omega-3 fatty acids or esters thereof wherein thefeed composition as a final mixture comprises about 0.01% to about 60%by weight of the algal composition and wherein the animal is a gestatingsow, a postpartum sow, another species of agricultural animal, acompanion animal, or a human, and increasing intestinal transport in theoffspring of the animal.

In accordance with this embodiment, the algal composition can be in theform of dried algae or an oil derived from the algae and the omega-3fatty acids can comprise C22 or C20 omega-3 fatty acids. Also inaccordance with this embodiment, the feed composition as a final mixturecan comprise about 0.01% to about 3.0% by weight, about 0.01% to about4.0% by weight, about 0.01% to about 1.5% by weight, about 0.01% toabout 1.0% by weight, about 0.01% to about 0.8% by weight, about 0.01%to about 0.5% by weight, about 0.01% to about 0.3% by weight, about 0.1%to about 0.5% by weight, about 0.01% to about 18% by weight, about 0.01%to about 20% by weight, about 0.01% to about 30% by weight, about 0.01%to about 40% by weight, about 0.01% to about 50% by weight, or about0.01% to about 60% by weight of the algal composition.

Also in accordance with this embodiment, the feed composition as a finalmixture can further comprise omega-6 fatty acids or esters thereof, thefeed composition can be administered during lactation, gestation, ordaily to the animal, the feed composition as a final mixture can furthercomprise an antioxidant, the omega-3 fatty acids in the feed compositioncan be stabilized by encapsulation, the omega-3 fatty acids can comprisedocosahexaenoic acid and eicosapentaenoic acid, and the omega-3 fattyacids can comprise docosahexaenoic acid, eicosapentaenoic acid, anddocosapentanoic acid. Further in accordance with this embodiment, theratio of docosahexaenoic acid to eicosapentaenoic acid can be about60:1, about 30:1, about 28:1, about 25:1, about 20:1, about 15:1, about10:1, about 5:1, or about 2:1 the species of agricultural animals can beselected from the group consisting of a chicken, a horse, a pony, a cow,a turkey, a pheasant, a quail, an ovine animal, a goat, an ostrich, anda duck, and the companion animal can be selected from the groupconsisting of a canine species and a feline species.

In yet another embodiment, a method of increasing intestinal transportof nutrients in a piglet is provided. The method comprises the steps ofadministering to the piglet a feed composition comprising an algalcomposition comprising omega-3 fatty acids or esters thereof wherein thealgal composition comprises docosahexaenoic acid and eicosapentaenoicacid and the docosahexaenoic acid to eicosapentaenoic acid ratio in thealgal composition is about 30:1 to about 1:1, and increasing intestinaltransport in the piglet.

In accordance with this embodiment, the algal composition can be in theform of dried algae or an oil derived from the algae, or residuals fromdried algae or algal oils, and the omega-3 fatty acids can comprise C22or C20 omega-3 fatty acids. Also in accordance with this embodiment, thefeed composition as a final mixture can comprise about 0.01% to about3.0% by weight, about 0.01% to about 4.0% by weight, about 0.01% toabout 1.5% by weight, about 0.01% to about 1.0% by weight, about 0.01%to about 0.8% by weight, about 0.01% to about 0.5% by weight, about0.01% to about 0.3% by weight, about 0.1% to about 0.5% by weight, about0.01% to about 18% by weight, about 0.01% to about 20% by weight, about0.01% to about 30% by weight, about 0.01% to about 40% by weight, about0.01% to about 50% by weight, or about 0.01% to about 60% by weight ofthe algal composition.

Also in accordance with this embodiment, the feed composition as a finalmixture can further comprise omega-6 fatty acids or esters thereof, thefeed composition can be administered daily to the animal, the feedcomposition as a final mixture can further comprise an antioxidant, theomega-3 fatty acids in the feed composition can be stabilized byencapsulation, and the omega-3 fatty acids can further comprisedocosapentanoic acid. Further in accordance with this embodiment, theratio of docosahexaenoic acid to eicosapentaenoic acid can be about30:1, about 28:1, about 25:1, about 20:1, about 15:1, about 10:1, about5:1, or about 1:1.

In still another embodiment, a method of increasing intestinal transportof nutrients in the offspring of an animal is provided. The methodcomprises the steps of administering to the animal a feed compositioncomprising a non-algal composition comprising omega-3 fatty acids oresters thereof wherein the docosahexaenoic acid to eicosapentaenoic acidratio in the non-algal composition is about 30:1 to about 1:1 andwherein the animal is a species of agricultural animal other than swine,a companion animal, or a human, and increasing intestinal transport inthe offspring of the animal.

In accordance with this embodiment, the omega-3 fatty acids can compriseC22 or C20 omega-3 fatty acids. Also in accordance with this embodiment,the feed composition as a final mixture can comprise about 0.01% toabout 3.0% by weight, about 0.01% to about 4.0% by weight, about 0.01%to about 1.5% by weight, about 0.01% to about 1.0% by weight, about0.01% to about 0.8% by weight, about 0.01% to about 0.5% by weight,about 0.01% to about 0.3% by weight, about 0.1% to about 0.5% by weight,about 0.01% to about 18% by weight, about 0.01% to about 20% by weight,about 0.01% to about 30% by weight, about 0.01% to about 40% by weight,about 0.01% to about 50% by weight, about 0.01% to about 60% by weight,about 0.01% to about 70% by weight of the algal composition.

Also in accordance with this embodiment, the feed composition as a finalmixture can further comprise omega-6 fatty acids or esters thereof, thefeed composition can be administered during lactation, gestation, ordaily to the animal, the feed composition as a final mixture can furthercomprise an antioxidant, the omega-3 fatty acids in the feed compositioncan be stabilized by encapsulation, and the omega-3 fatty acids canfurther comprise docosapentanoic acid. Further in accordance with thisembodiment, the ratio of docosahexaenoic acid to eicosapentaenoic acidcan be about 25:1, about 20:1, about 15:1, about 10:1, about 5:1, orabout 2:1, the species of agricultural animals can be selected from thegroup consisting of a chicken, a horse, a pony, a cow, a turkey, apheasant, a quail, an ovine animal, a goat, an ostrich, and a duck, andthe companion animal can be selected from the group consisting of acanine species and a feline species.

In another illustrative embodiment, a method of increasing the growthperformance of an offspring of an animal is provided. The methodcomprises the steps of administering to the animal a feed compositioncomprising an algal composition comprising omega-3 fatty acids or estersthereof wherein the algal composition comprises docosahexaenoic acid andeicosapentaenoic acid and the docosahexaenoic acid to eicosapentaenoicacid ratio in the algal composition is about 60:1 to about 1:1 andwherein the animal is a gestating sow, a postpartum sow, another speciesof agricultural animal, a companion animal, or a human, and increasingthe growth performance of the offspring of the animal. In anotherembodiment, the growth performance is selected from a group consistingof an increased growth rate of the offspring, a reduced feed to weightgain ratio for the offspring, and an increase in the efficiency of feedutilization.

In another aspect, a method is provided of increasing the growthperformance of an offspring of an animal. The method comprises the stepsof administering to the animal a feed composition comprising an algalcomposition comprising omega-3 fatty acids or esters thereof wherein thefeed composition as a final mixture comprises about 0.01% to about 60%by weight of the algal composition and wherein the animal is a gestatingsow, a postpartum sow, another species of agricultural animal, acompanion animal, or a human, and increasing the growth performance ofthe offspring of the animal. In one aspect, the growth performance isselected from a group consisting of an increased growth rate of theoffspring and a reduced feed to weight gain ratio for the offspring.

In yet another embodiment, a method is provided of increasing the growthperformance of an offspring of an animal. The method comprises the stepsof administering to the animal a feed composition comprising a non-algalcomposition comprising omega-3 fatty acids or esters thereof wherein thedocosahexaenoic acid to eicosapentaenoic acid ratio in the non-algalcomposition is about 30:1 to about 1:1 and wherein the animal is aspecies of agricultural animal other than swine, a companion animal, ora human, and increasing the growth performance of the offspring of theanimal. In this embodiment, the growth performance can be selected froma group consisting of an increased growth rate of the offspring and areduced feed to weight gain ratio for the offspring.

In still another embodiment, a method is provided of increasing thegrowth performance of an offspring of an animal. The method comprisesthe steps of administering to the animal a feed composition comprising anon-algal composition comprising omega-3 fatty acids or esters thereof,wherein the feed composition as a final mixture can comprise about 0.01%to about 90% by weight of the non-algal composition, wherein the animalis a species of agricultural animal, a companion animal, or a human, andincreasing the growth performance of the offspring of the animal. Inthis embodiment, the growth performance can be selected from a groupconsisting of an increased growth rate of the offspring and a reducedfeed to weight gain ratio for the offspring.

In another embodiment, a method is provided of increasing intestinaltransport in an offspring of a swine. The method comprises the steps ofadministering to the swine a feed composition comprising a non-algalcomposition comprising omega-3 fatty acids or esters thereof wherein thedocosahexaenoic acid to eicosapentaenoic acid ratio in the non-algalcomposition is about 30:1 to about 2:1.

In yet another embodiment, a method is provided of increasing the growthperformance in an offspring of a swine. The method comprises the stepsof administering to the swine a feed composition comprising a non-algalcomposition comprising omega-3 fatty acids or esters thereof wherein thedocosahexaenoic acid to eicosapentaenoic acid ratio in the non-algalcomposition is about 30:1 to about 2:1.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows active glucose (panel A) and glutamine (panel B) transportin jejunum samples obtained from piglets weaned at 14-17 days of agefrom sows fed one of four diets: (1) a basal corn/soybean meal control(no added fat, CONT); (2) the basal diet supplemented with protectedfish oil (PFO); (3) the basal diet supplemented with DHA fats fromSchizochytrium algae (a1DHA); or (4) the basal diet supplemented withdried coconut fat (COCO). Piglets (n=4 per treatment) were deprived offeed for 24 hours post-weaning. Means without a common letter differ,P<0.05. Pooled SEM are shown.

FIG. 2 shows GLUT2 protein expression in jejunum samples obtained frompiglets weaned at 14-17 days of age. The piglets were from sows fed: (1)a basal corn/soybean meal control (no added fat, CONT); (2) the basaldiet supplemented with protected fish oil (PFO); (3) the basal dietsupplemented with DHA fats from Schizochytrium algae (a1DHA); or (4) thebasal diet supplemented with dried coconut fat (COCO). Piglets (n=4 pertreatment) were deprived of feed for 24 hours post-weaning and pooledSEM are shown.

FIG. 3 shows SGLT1 protein expression in jejunum samples obtained frompiglets weaned at 14-17 days of age. The sows were fed: (1) a basalcorn/soybean meal control (no added fat, CONT); (2) the basal dietsupplemented with protected fish oil (PFO); (3) the basal dietsupplemented with DHA fats from Schizochytrium algae (a1DHA); or (4) thebasal diet supplemented with dried coconut fat (COCO). Piglets (n=4 pertreatment) were deprived of feed for 24 hours post-weaning. Meanswithout a common letter differ, P<0.05. Pooled SEM are shown.

FIG. 4 shows glucose, glycogen and total glycosyl concentrations inlongissimus muscle (panel A) and liver (panel B) samples obtained frompiglets weaned at 14-17 days of age. The piglets were from sows fed: (1)a basal corn/soybean meal control (no added fat, CONT); (2) the basaldiet supplemented with protected fish oil (PFO); (3) the basal dietsupplemented with DHA fats from Schizochytrium algae (a1DHA); or (4) thebasal diet supplemented with dried coconut fat (COCO). Piglets (n=4 pertreatment) were deprived of feed for 24 hours post-weaning. Differentletters represent significant differences (P<0.05). Pooled SEM areshown.

FIG. 5 shows ex vivo active glucose uptake by proximal jejunum ofpiglets at 21 days of age after deprivation of feed for 24 hours tosimulate weaning stress. Dams were fed the control (Cont) and protectedfish oil (PFO) dietary regimens during gestation and/or lactation (G/L).Data represent the means of 6 piglets per treatment. Means without acommon letter are significantly different (P<0.05).

FIG. 6 shows an abundance of GLUT2 protein in the crude brush bordermembranes (BBM) (panel A) and total tissue preparations (panel B) fromthe proximal jejunum of piglets at 21 days of age after deprivation offeed for 24 hours to simulate weaning stress. Dams were fed the control(Cont) and protected fish oil (PFO) dietary regimens during gestationand/or lactation (G/L). Data represent the means SE of 6 piglets pertreatment. Means without a common letter are significantly different(P<0.05).

FIG. 7 shows an abundance of SGLT1 protein in the crude brush bordermembranes (BBM) (panel A) and total tissue preparations (panel B) fromthe proximal jejunum of piglets at 21 days of age after deprivation offeed for 24 hours to simulate weaning stress. Dams were fed the control(Cont) and protected fish oil (PFO) dietary regimens during gestationand(or) lactation (G/L). Data represent the means±SE of 6 piglets pertreatment. Means without a common letter are significantly different(P<0.05), and the Cont/Cont and Cont/PFO differed at P<0.10.

FIG. 8 shows jejunum glucose uptake in chicks. Values are least squaresmeans±SEM (n=10/treatment). Different letters represent significantdifferences at P<0.05.

FIG. 9 shows jejunum glutamine uptake in chicks. Values are leastsquares means±SEM (n=10/treatment).

DETAILED DESCRIPTION OF THE INVENTION

While the invention is susceptible to various modifications andalternative forms, specific embodiments will herein be described indetail. It should be understood, however, that there is no intent tolimit the invention to the particular forms described, but on thecontrary, the intention is to cover all modifications, equivalents, andalternatives falling within the spirit and scope of the invention.

Methods and compositions for increasing intestinal transport ofnutrients in an offspring of an animal are described. More particularly,methods and compositions are described for administration to an animalof a feed composition supplemented with a composition comprising omega-3fatty acids or esters thereof, to increase the intestinal transport ofnutrients in the offspring of the animal. For example, the methods andcompositions described herein may increase intestinal transport ofnutrients including, but not limited to, vitamins, lipids, minerals,amino acids (e.g., glutamine, etc.), carbohydrates (e.g., glucose,etc.), proteins, and the like. Additionally, the methods andcompositions described are useful to increase tissue energy stores(e.g., muscle glycogen units or muscle glycosyl units).

The compositions described herein contain, in particular, a source ofomega-3 fatty acids or esters thereof, such as products from algalsources (e.g., algal oils, dried algal products, and residuals andderivatives thereof), non-algal sources (e.g., oils, dried products, andderivatives of non-algal marine sources, as well as nuts, seeds, andplant products), or combinations thereof. The algal and non-algalproducts serve as a source of omega-3 fatty acids/esters and theirmetabolites, such as eicosapentaenoic acid (EPA), docosahexaenoic acid(DHA), and docosapentaenoic acid (DPA), or mixtures thereof.

Any source of omega-3 fatty acids may be used in the methods andcompositions described herein. In one embodiment, omega-3 fatty acidsources useful in the methods and compositions described herein compriseC₂₂ omega-3 fatty acids and/or C₂₀ omega-3 fatty acids. In anotherembodiment, compositions for use as a source of omega-3 fatty acids inthe feed composition as a final mixture will have a DHA:EPA ratio Instill another illustrative embodiment, the feed composition as a finalmixture as described herein comprises DHA and EPA in a DHA:EPA ratio ofabout 1:1 to about 60:1. In another illustrative embodiment, the finalfeed compositions as described herein comprise DHA and EPA in a ratio offrom about 8:1 to about 30:1, or about 30:1, about 28:1, about 25:1,about 20:1, about 15:1, about 10:1, about 5:1, or about 2:1.

Fatty acids with no double bonds are termed saturated fatty acids, thosewith one double bond are termed monounsaturated fatty acids, and thosewith multiple double bonds are termed polyunsaturated fatty acids.Overall digestibility appears to increase with the degree ofunsaturation. A convenient shorthand system is used in thisspecification to denote the structure of fatty acids. This system uses anumber denoting the number of carbons in the hydrocarbon chain, followedby a colon and a number indicating the number of double bonds in themolecule, and then by a “w6” or a “w3” to denote “omega-6” or “omega-3”,respectively (e.g., 22:5w6). The “w6” or a “w3” denotes the location ofthe first double bond from the methyl end of the fatty acid molecule.Trivial names in the w6 series of fatty acids include linoleic acid(18:2w6), gamma-linoleic acid (18:3w6), and arachidonic acid (20:4w6).The only fatty acid in the w3 series with a trivial name isalpha-linolenic acid (18:3w3). For the purposes of this application afatty acid with the nomenclature 20:5w3 is eicosapentaenoic acid, withthe nomenclature 22:6w3 is docosahexaneoic acid, and with thenomenclature 22:5w3 is docosapentaenoic acid.

The methods of the present invention utilize an omega-3 fattyacid-containing composition as a source of long chain omega-3 fattyacids, such as eicosapentaenoic acid, docosahexaneoic acid,docosapentaenoic acid, and esters thereof, to increase the intestinaltransport of nutrients in an offspring of an animal. The omega-3 fattyacid-containing composition used herein can be obtained from an algalsource or a non-algal source. In one embodiment, the feed composition issupplemented with an omega-3 fatty acid-containing compositioncomprising DHA and EPA, wherein the DHA:EPA ratio in the feedcomposition as a final mixture is about 1:1 to about 30:1. In oneaspect, this feed composition can be fed to piglets. In anotherembodiment, the DHA:EPA ratio in the final feed composition is about30:1 to about 2:1. In one illustrative embodiment, the feed compositioncan contain a non-algal source of omega-3 fatty acids. In yet anotherembodiment, the DHA:EPA ratio in the final feed composition is about28:1, about 25:1, about 20:1, about 15:1, about 10:1, about 5:1, orabout 2:1.

A biologically effective amount of the omega-3 fatty acid-containingcomposition can be administered to increase the intestinal transport ofnutrients in the offspring of animals. By “biologically effectiveamount” is meant an amount of the omega-3 fatty acid-containingcomposition capable of increasing the intestinal transport of nutrientsin the offspring of an animal by any mechanism, including thosedescribed herein. Additionally, a biologically effective amount of theomega-3 fatty acid-containing composition can be an amount capable ofincreasing tissue energy stores, and/or an amount effective to increasegrowth performance (e.g., increasing growth rate, reducing the feed toweight gain ratio, increasing weaning weight, or increasing theefficiency of feed utilization).

In one illustrative embodiment, the feed compositions of the inventionthat contain omega-3 fatty acids are administered to the animals orally,but any other effective method of administration known to those skilledin the art may be utilized.

In illustrative embodiments, the feed composition as a final mixture maycomprise an algal derived composition or a non-algal derivedcomposition, such as a non-algal marine product (e.g., fish oil or fishmeal), or a nut, seed, or plant derived product (e.g., walnut, flaxseed,canola, soybean oil, or corn oil, or a derivative thereof), orcombinations thereof. In illustrative embodiments, the feed compositionas a final mixture may be supplemented with any omega-3 fattyacid-containing composition, and may include, for example, an algal oil,a dried algal product (including dried whole cells and ground algalproducts), a fish oil (e.g., salmon oil or another fish oil from a NorthAtlantic cold water fish), fish meal, or an oil derived from fish meal,or a mixture thereof, or residuals from any of these sources of omega-3fatty acids to provide a source of omega-3 fatty acids/esters in amixture with an art-recognized animal feed blend.

In one illustrative aspect, the feed composition as a final mixture maybe administered to the animal for any time period that is effective toincrease the intestinal transport of nutrients in the offspring of theanimal. For example, the feed composition may be fed to a female animaldaily for the lifetime of the animal. Alternatively, the feedcomposition may be administered to the animal for a shorter time period.In one embodiment, the feed composition can be administered to agestating sow, a postpartum sow, a piglet, another species ofagricultural animal, a companion animal, or a human (e.g., to increasethe longevity of the human or the animal). Illustratively, the companionanimal, the human, or the species of agricultural animal may be agestating, a lactating, or a postpartum animal.

In another embodiment, the feed composition is administered to apostnatal animal, including a nursing animal or an animal being weanedor an animal after weaning. In another embodiment, the feed compositioncan be administered to a prenatal animal in utero by feeding thecomposition to a gestating mother. In yet another embodiment, the feedcomposition can be administered to a nursing animal by feeding thecomposition to a lactating mother, or alternatively, by feeding thecomposition directly to the nursing animal through a prepared diet(e.g., a formula). The time periods for administration of the feedcomposition described above are nonlimiting and it should be appreciatedthat any time period determined to be effective to increase theintestinal transport of nutrients in the offspring of the animal may beused.

In one embodiment, as described herein, a species of agricultural animalother than a pig may be fed the feed composition and those species mayinclude bovine species (e.g., cattle and bison), equine species (e.g.,horses, ponies, and donkeys), ovine species (e.g., sheep), caprinespecies (e.g., goats), rabbits, and poultry (e.g., chickens, turkeys,pheasant, ducks, ostrich, emu, quail, and geese). As used herein, aspecies of agricultural animal other than a pig may include any animalthat is raised for personal use (e.g., for providing food, fuel, etc.)or for profit. In yet another embodiment, a companion animal may be fedthe compositions described herein and a companion animal include anyanimal that is kept or raised for companionship purposes, for example,canine and feline species.

In various illustrative embodiments, any animal feed blend known in theart may be used to make the feed composition such as rapeseed meal,flaxseed meal, cottonseed meal, soybean meal, and cornmeal. The animalfeed blend can be supplemented with an omega-3 fatty acid-containingcomposition, but other ingredients may optionally be added to the animalfeed blend. Optional ingredients of the animal feed blend include sugarsand complex carbohydrates such as both water-soluble and water-insolublemonosaccharides, disaccharides and polysaccharides. Optional amino acidingredients that may be added to the feed blend are arginine, histidine,isoleucine, leucine, lysine, methionine, phenylalanine, threonine,tryptophan, valine, tyrosine ethyl HCl, alanine, aspartic acid, sodiumglutamate, glycine, proline, serine, cysteine ethyl HCl, and analogs,and salts thereof. Vitamins that may be optionally added are thiamineHCl, riboflavin, pyridoxine HCl, niacin, niacinamide, inositol, cholinechloride, calcium pantothenate, biotin, folic acid, ascorbic acid, andvitamins A, B, K, D, E, and the like. Optional lipid blends of animal orplant origin or fiberous ingredients could also be added. Proteiningredients may also be added and include protein obtained from meatmeal or fish meal, liquid or powdered egg, fish solubles, and the like.Any medicament ingredients known in the art may also be added to theanimal feed blend such as antibiotics.

In an illustrative embodiment, antioxidants may be added to the feedcomposition to prevent oxidation of the fatty acids present in theomega-3 fatty acid-containing composition used to supplement the feedcomposition, such as the omega-3 long chain fatty acids,eicosapentaenoic acid, docosahexaneoic acid, and docosapentaenoic acid.Oxidation of fatty acids occurs over time and may be affected by suchconditions as moisture and the presence of mineral catalysts and by suchcharacteristics of fatty acids as the number of double bonds andpositioning and configuration of bonds. Oxidation of these omega-3 fattyacids can be prevented by the introduction of naturally-occurringantioxidants, such as beta-carotene, vitamin E, vitamin C, andtocopherol or of synthetic antioxidants such as butylatedhydroxytoluene, butylated hydroxyanisole, tertiary-butylhydroquinone,propyl gallate or ethoxyquin to the feed composition. Compounds whichact synergistically with antioxidants can also be added such as ascorbicacid, citric acid, and phosphoric acid. The amount of antioxidantsincorporated in this manner depends on requirements such as productformulation, shipping conditions (e.g., shipping under a nitrogenblanket), packaging methods, and desired shelf life.

In one embodiment, the omega-3 fatty acid-containing composition used tosupplement the feed composition is derived from a high purity algalpreparation that comprises a high content of DHA. In one aspect, thealgal preparation may comprise a high DHA:EPA ratio, i.e., the amount ofDHA in the composition can be greater than or equal to the amount of EPAin the composition (e.g., up to a 60:1 ratio of DHA:EPA). In analternative embodiment, no EPA is present in the algal composition.Various ratio embodiments are described herein. In one embodiment, thefeed composition as a final mixture can be supplemented with an omega-3fatty acid-containing composition derived from algae, such as oils,gels, pastes, dried products, and residuals or derivatives thereof. Inother embodiments, the omega-3 fatty acid-containing composition mayinclude whole algal cell products, ground algal products, or residualproducts remaining from the production of oils, gels, pastes, and driedproducts, or derivatives thereof. In illustrative aspects, the algalproduct may be obtained from any algal source, including marine orfreshwater algal sources.

In various embodiments, the algal sources may include, but are notlimited to, species of Schizochytrium, Spirulina, Chlorella,Chaetoceros, Cyclotella, Isochrysis, Nonnocholoropsis, Nitzschia,Phyaeodactylum, as well as dinoflagellates, including species ofAmphidinium, Ceratium, Cochlodinium, Crypthecodinium (e.g.,Crypthecodinium cohnii), Gonyaulax, and Peridinium. In anotherembodiment, the omega-3 fatty acid-containing composition derived fromalgae may include a composition derived from a genetically modifiedorganisms, modified by expression of an algal gene. Any non-toxic algalsource capable of increasing intestinal transport of nutrients in ananimal may be used. In various embodiments, algal products as hereindescribed provide a source of omega-3 polyunsaturated long chain fattyacids including eicosapentaenoic acid (20:5w3), docosahexaneoic acid(22:6w3), and docosapentaenoic acid (22:5w3). In various illustrativeembodiments, the omega-3 fatty acid-containing composition derived fromalgae has a DHA:EPA ratio ≧1:1, ≧5:1, ≧10:1, ≧15:1, ≧20:1, or ≧25:1.

In another embodiment, the animal feed blend is supplemented with anomega-3 fatty acid-containing composition derived from a non-algalsource, such as fish oils or fish meal, as well as plant, nut, or seedoils, or a derivative thereof, or a combination thereof. The omega-3fatty acid-containing composition derived from a non-algal source mayalso include compositions derived from a genetically modified organism.For example, genetically modified plants, including transgenic plants,may be used as a non-algal source of omega-3 fatty acids. In addition,nutritionally enhanced plants or seeds may be used as a non-algal sourceof omega-3 fatty acids. Examples of plants that may be geneticallymodified or nutritionally enhanced for use as a non-algal sourceinclude, but are not limited to, corn, canola, soybean, flax, rapeseed,and hominy.

The non-algal oils described herein may be obtained from any source. Inone embodiment, the non-algal oil source is North Atlantic cold waterfish. Fish oils provide a source of both omega-3 and omega-6 fattyacids, but are a particularly good source of omega-3 polyunsaturatedfatty acids. The omega-3 polyunsaturated long chain fatty acidseicosapentaenoic acid (20:5w3), docosahexaneoic acid (22:6w3), anddocosapentaenoic acid (22:5w3) are typical of fish oils and togethercomprise about 25-38% by weight of the fish oil. Omega-6 polyunsaturatedfatty acids present in fish oils include linoleic acid and arachidonicacid and are present at lesser concentrations of about 10% by weight.

Oils are understood to be lipids or fats including the glyceride estersof fatty acids along with associated phosphatides, sterols, alcohols,hydrocarbons, ketones, alkyl esters, salts, and related compounds.Further, as described herein, dried products include algal and non-algalproducts prepared by any method known in the art, and may includespray-dried or freeze-dried products. The algal and non-algalcompositions described herein may include whole cell products, groundproducts, or residuals or derivatives thereof.

In various illustrative aspects, the oils or fatty acid ester componentsmay be added in an unprocessed form or in pure form, or may beconjugated or unconjugated, including supplements (e.g., conjugatedlinoleic acids). Illustratively, the fatty acid esters added to the feedcomposition may be triglycerides, diglycerides, monoglycerides,phospholipids, lysopholipids, or can be chemically beneficiated orenzymatically beneficiated for enhanced content of desired fatty acidesters.

In one embodiment, the feed compositions described herein may alsocomprise omega-6 fatty acids or esters thereof, as described in U.S.Pat. No. 7,084,175 and U.S. patent application Ser. No. 10/142,685,incorporated herein by reference. Illustratively, the omega-6 fattyacids usable in the present invention can be unsaturated fatty acidshaving at least two carbon-carbon double bonds such as 2,4-decadienoicacid, linolenic acid, gamma-linolenic acid, 8,10,12-octadecatrienoicacid and arachidonic acid. In another embodiment, the omega-6 fatty acidcan be gamma-linolenic acid. In other embodiments, omega-6 fattyacids/esters for use in the feed composition of the present inventioninclude omega-6 fatty acids/esters derived from an art-recognized mealsuch as corn meal or soybean meal or from oils such as corn oil,cottonseed oil, soybean oil, safflower oil, sunflower oil, linseed oil,borage oil, blackcurrant oil, evening primrose oil, and the like.

In one illustrative aspect, the feed composition described herein issupplemented with concentrations of an omega-3 fatty acid-containingcomposition, such as algal oil, gel, paste, dried products, or acombination thereof, or residuals thereof, sufficient to provide amountsof omega-3 fatty acids/esters in the feed composition as a final mixturethat are effective in increasing the intestinal transport of nutrientsin the offspring of an animal. Alternatively, the feed composition maybe supplemented with an omega-3 fatty acid-containing composition, suchas fish oil, fish meal, plant-derived products, or combinations thereof,sufficient to provide amounts of omega-3 fatty acids/esters in the feedcomposition as a final mixture that are effective in increasing theintestinal transport of nutrients in the offspring of an animal. Inanother embodiment, the feed composition may be supplemented with acombination of algal and non-algal omega-3 fatty acid-containingsources.

In one illustrative embodiment, the feed composition can be supplementedwith an omega-3 fatty acid-containing composition in an amount of about0.01% to about 60% by weight of the feed composition as a final mixture.In another embodiment the feed composition can be supplemented with anomega-3 fatty acid-containing composition in an amount of about 0.1% toabout 0.5% by weight of the feed composition as a final mixture. In yetanother embodiment, the feed composition can be supplemented with anomega-3 fatty acid-containing composition in an amount of about 0.01% toabout 0.3% by weight, about 0.01% to about 0.5% by weight, about 0.01%to about 0.8% by weight, about 0.01% to about 1.0% by weight, about0.01% to about 1.5% by weight, about 0.01% to about 3.0% by weight,about 0.01% to about 4.0% by weight, about 0.01% to about 18% by weight,about 0.01% to about 20% by weight, about 0.01% to about 30% by weight,about 0.01% to about 40% by weight, about 0.01% to about 50% by weight,about 0.01% to about 60% by weight, about 0.01% to about 70% by weight,or about 0.01% to about 90% by weight of the feed composition as a finalmixture.

In each of these embodiments it is to be understood that the percentageof the omega-3 fatty acid-containing composition by weight of the feedcomposition refers to the final feed composition (i.e., the feedcomposition as a final mixture) containing the animal feed blend, theomega-3 fatty acid-containing composition (e.g., algal oil, ground algaeor other dry algal product, or residuals thereof, or fish oil, etc.),and optionally added ingredients. In such embodiments of the invention,the omega-3 fatty acid-containing composition may be derived from anytype of algal or non-algal source.

In various aspects, the omega-3 fatty acid-containing composition asdescribed herein may be administered in an unencapsulated or anencapsulated form in a mixture with an animal feed blend. Encapsulationprotects the omega-3 fatty acids/esters and omega-6 fatty acids/estersfrom breakdown and/or oxidation prior to digestion and absorption of thefatty acids/esters by the animal (i.e., encapsulation increases thestability of fatty acids) and provides a dry product for easier mixingwith an animal feed blend. The omega-3 fatty acids/esters and omega-6fatty acids/esters can be protected in this manner, for example, bycoating the oil with a protein or any other substances known in the artto be effective encapsulating agents such as polymers, waxes, fats, andhydrogenated vegetable oils. For example, an oil or other algal ornon-algal product, may be encapsulated using an art-recognized techniquesuch as a Na²⁺-alginate encapsulation technique wherein the oil iscoated with Na²⁺-alginate followed by conversion to Ca²⁺-alginate in thepresence of Ca²⁺ ions for encapsulation. Alternatively, the oil or otheralgal or non-algal product may be encapsulated by an art-recognizedtechnique such as enrobing the fatty acids to stabilize the fatty acidsor prilling (i.e., atomizing a molten liquid and cooling the droplets toform a bead). For example, the oil or other algal or non-algal productmay be prilled in hydrogenated cottonseed flakes or hydrogenated soybean oil to produce a dry oil. In various embodiments, the oil or otheralgal or non-algal product may be used in an entirely unencapsulatedform, an entirely encapsulated form, or mixtures of unencapsulated andencapsulated oil may be added to the feed composition.

In various embodiments, the feed compositions described herein, when fedto in utero through the mother (e.g., a gestating sow) and/or topostnatal animals (e.g., by lactation through a postpartum sow ordirectly to the postnatal animal), may result not only in increases inintestinal transport, but also in benefits regarding insulinsensitivity, and in increases in growth performance of the postnatalanimals (e.g., a piglet). Any of the embodiments described above can beused to increase the growth performance (e.g., increased growth rate,reduced feed to weight gain ratio, or increased efficiency of feedutilization) of the offspring of an animal.

Accordingly, in one embodiment, a method for increasing growthperformance of the postnatal animal is provided. The method comprisesthe step of administering to the postnatal animal or the mother a feedcomposition comprising an algal composition comprising omega-3 fattyacids or esters thereof wherein the feed composition as a final mixturecomprises about 0.01% to about 60% by weight of the algal compositionand wherein the animal is a sow, a piglet, another species ofagricultural animal, a companion animal, or a human.

In accordance with this embodiment, the algal composition can be in theform of dried algae or an oil derived from the algae, or a residual ofeither composition, and the omega-3 fatty acids can comprise C₂₂ or C₂₀omega-3 fatty acids. Also in accordance with this embodiment, the feedcomposition as a final mixture can comprise about 0.01% to about 3.0% byweight, about 0.01% to about 4.0% by weight, about 0.01% to about 1.5%by weight, about 0.01% to about 1.0% by weight, about 0.01% to about0.8% by weight, about 0.01% to about 0.5% by weight, about 0.01% toabout 0.3% by weight, about 0.1% to about 0.5% by weight, about 0.01% toabout 18% by weight, about 0.01% to about 20% by weight, about 0.01% toabout 30% by weight, about 0.01% to about 40% by weight, about 0.01% toabout 50% by weight, or about 0.01% to about 60% by weight of the algalcomposition.

Also in accordance with this embodiment, the feed composition as a finalmixture can further comprise omega-6 fatty acids or esters thereof, thefeed composition can be administered during lactation, or daily to theanimal, the feed composition as a final mixture can further comprise anantioxidant, the omega-3 fatty acids in the feed composition can bestabilized by encapsulation, the omega-3 fatty acids can comprisedocosahexaenoic acid and eicosapentaenoic acid, and the omega-3 fattyacids can comprise docosahexaenoic acid, eicosapentaenoic acid, anddocosapentanoic acid. Further in accordance with this embodiment, theratio of docosahexaenoic acid to eicosapentaenoic acid can be about60:1, about 30:1, about 25:1, about 20:1, about 15:1, about 10:1, about5:1, about 2:1, or about 1:1, the species of agricultural animals can beselected from the group consisting of a chicken, a horse, a pony, a cow,a turkey, a pheasant, a quail, an ovine animal, a goat, an ostrich, anda duck, and the companion animal can be selected from the groupconsisting of a canine species and a feline species.

This embodiment of the invention can also comprise method embodimentswhere the postnatal animal is fed a feed composition supplemented with anon-algal source of omega-3 fatty acids under any of the conditionsdescribed above where the DHA:EPA ratio is about 30:1 to about 1:1.

While certain embodiments of the present invention have been describedand/or exemplified below, it is contemplated that considerable variationand modification thereof are possible. Accordingly, the presentinvention is not limited to the particular embodiments described and/orexemplified herein.

EXAMPLE 1 Animals and Experimental Design

Twenty dams (Ausgene Line 20 dams×SPI sixes) were fed one of fourexperimental diets for approximately 150 days prior to farrowing (Table1). The four experimental dietary treatments consisted of (1) basalcorn/soybean meal control (no added fat, “CONT”); (2) the basal dietsupplemented with protected fish oil (FERTILIUM™ or GROMEGA365™; JBSUnited Inc., Sheridan, Ind. [i.e., PFO]); (3) the basal dietsupplemented with DHA fats from Schizochytrium algae (a1DHA); (4) thebasal diet supplemented with dried coconut fat (COCO).

The fatty acid profiles of the diets are presented in Table 2. Both theprotected fish oil and a1DHA ingredients had high (n-3) PUFAconcentrations and contained 29 and 43% total fat, respectively. Therest of these ingredients comprised of protein and carbohydrate. a1DHAcontained 40% DHA and 2% EPA by percentage of fat, while PFO hadapproximately 13% EPA and 13% DHA. The total fat of the four dietsdiffered. However, the DHA percentage of the a1DHA diet was equal to theDHA percentage in the PFO diet. The raw COCO fat ingredient wascomprised of 88% saturated fat (high in saturated fatty acidsC10:0-C16:0) as a percentage of fat. All diets (Table 1) met andexceeded the nutrient requirements for gestating and lactating sows [seeNCR, Nutritional Requirements of Swine, 10^(th) ed., Washington, D.C.:Natl. Acad. Press (1998)] and all piglets had access to water at alltimes.

While farrowing in two replicate groups over two weeks, litters werestandardized to ten piglets per litter within 24 hours of birth, withcross fostering only occurring within treatment. Subsequently, at 14-17days of age, one medium size piglet (4.1 kg±0.49) per litter wasrandomly separated from the dam, grouped penned with piglets from otherlitters, and fasted overnight to simulate the weaning process (total n=4per treatment). Following the simulated weaning, piglets were killed bycaptive bolt gun followed by immediate exsanguination and tissuesexcised. Small intestinal jejunum, liver and muscles samples werecollected and snap frozen in liquid nitrogen and jejunum samples placedin 10% formalin for subsequent analysis.

EXAMPLE 2 Fatty Acid Analysis

One week post farrowing, approximately 40 mL of mid-lactation milk wasobtained from four sows of each dietary treatment following an i.v.injection of 10 IU of oxytocin-S (Intervet, Millsboro, Del. USA) toinduce milk secretion. At random, several udders from each sow weremilked, pooled together and snap frozen on dry ice pending fatty acidanalysis. Lipids from milk, muscle and liver samples were extracted bythe method of Lepage and Roy [J. Lipid Res., 27: 114-120 (1986),incorporated herein by reference] with minor modifications. Briefly, 0.5g of tissue or 300 μL of milk were homogenized in 2.5 mL 4:1methanol:hexane and 200 μL of 3.7 mmol heptadecanoic acid/L methanol wasadded to each sample as an internal standard.

Fatty acid methyl esters were analyzed by gas chromatography on aHewlett-Packard model 6890 (Hewlett-Packard, Palo Alto, Calif.) fittedwith a Omegawax 320 (30 m×0.32 mm ID, 0.25 μm) capillary column(Sigma-Aldrich, St Louis, Mo. USA). Hydrogen was the carrier gas. Thetemperature program ranged from 80° C. to 250° C. with a temperaturerise of 5° C./min. The injector and detector temperatures were 250° C.and 1 μL of sample was injected and run splitless. Fatty acids wereidentified by their retention times on the column with respect toappropriate standards.

Feeding gestating and lactating dams the CONT, PFO, a1DHA or COCO diets,which varied considerably in their fatty acid profiles, changed thefatty acid composition of the milk accordingly (Table 3). The PFO anda1DHA diets had the highest (n-3) fatty acid proportions, thusreflecting each particular ingredient's DHA and EPA concentration. As aresult, milk from dams fed the CONT and COCO diets had higher(n-6):(n-3) fatty acid ratios than that from dams fed the other twodiets. Additionally, milk from the sows fed the COCO diet was higher intotal saturated fatty acids compared to the PFO and a1DHA, but notcompared to the CONT milk (P<0.05, Table 3). Of the saturated fattyacids found in the sow's milk, C12:0 was six-fold higher in the COCOcompared to the CONT or (n-3) PUFA milk samples (P<0.05, Table 3).

Piglet small intestine and muscle fatty acids profiles are outlined inTable 4. The (n-3) PUFA fatty acid supplementation via the PFO maternaldiet increased piglet small intestine and muscle total (n-3) PUFAconcentration, 200 and 400%, respectively, vs. the CONT group (Table 4).This increase reflected the DHA and(or) EPA percentage of the sows'diets (Table2) and milk (Table 3), and corresponded significantly withdecreased (n-6):(n-3) ratios in all tissues tested (Table 4).

EXAMPLE 3 Ussing Chamber

Proximal jejunum samples starting 40 cm from the stomach consisting of a20-30 cm segment of proximal jejunum were removed and placed in chilledKrebs-Henseleit buffer (consisting in mmol/L: 25 NaHCO₃, 120 NaCl, 1MgSO₄, 6.3 KCl, 2 CaCl, 0.32 NaH₂PO₄; pH 7.4) for transport back to thelaboratory (<40 min) under constant aeration until clamped in the Ussingchambers. Two jejunal segments per pig were then stripped of outermuscle layers and immediately mounted in Ussing Chambers (DVC 1000 WorldPrecision Instruments, New haven, Conn. USA).

Each segment was bathed on its mucosal and serosal surfaces (openingarea 1.0 cm²) with 8 mL Krebs solution and gassed with 95% O₂-5% CO₂mixture. The intestinal segments were voltage clamped at zero mV by anexternal current after correction for solution resistance. After a 30minute period to allow the tissues to stabilize, they were challengedindependently with 10 mmol/L D-Glucose and 10 mmol/L L-glutamine whichwas added to serosal buffer, with equimolar (10 mmol/L) mannitol addedon the mucosal side.

Potential difference across the tissue was measured for 30 min aftereach challenge by open circuit conditions every 10 seconds due to ashort-circuit current being delivered by a voltage clamp apparatus. Thechange in maximal current was recorded and the tissue conductance wascalculated from the short-circuit current and potential difference usingohm's law. This was repeated on four different days with a total of fourpigs per treatment.

Ex-vivo jejunal nutrient absorption following the addition of 10 mmol/LD-glucose or L-glutamine was evaluated in three ways: (1) short circuitcurrent, which measures change in active ion transport; (2) conductance,which measures changes in total ion transport; and (3) passive iontransport, which is measured by changes in resistance.

Active transport was significantly greater following the addition ofD-glucose in tissue obtained from piglets of dams fed the a1DHA and PFOdiets vs. CONT piglets (P<0.05, FIG. 1, panel A). However, only thea1DHA treatment glucose transport significantly higher than in the COCOpiglets (P<0.05, FIG. 1, panel A). Compared with CONT tissues, activeD-glucose uptake of tissue from a1DHA piglets was 470% higher, but thePFO and COCO diets also resulted in greater uptake vs. the CONT (320%and 40%, respectively). See also FIG. 5. However, active L-glutamineuptake was only higher in tissue from piglets of dams fed the a1DHA dietas compared with CONT and PFO piglets (FIG. 1, panel B). Neither totalnor passive ion transport was affected by (n-3) PUFA or COCO dietarysupplementation (data not shown)

EXAMPLE 4 Determination of Muscle and Liver Glycogen

Samples of muscle (longissimus dorsi) or liver tissue (0.5 g) wereextracted in ice cold perchloric acid (0.5 mol/L) using a Tissue Tearorhomogenizer. Duplicate samples (300 μL) of each homogenate were thenprepared for glycogen hydrolysis with 0.3 g/L amyloglucosidase(Sigma-Aldrich, St Louis, Mo. USA) for 120 minutes at 38° C. Theincubation was stopped by the addition of 0.6 mol/L perchloric acid andthe samples clarified by centrifugation (1,500×g, 15 minutes at 4° C.).Glucose (HK) assay kits (Sigma-Aldrich, St Louis, Mo. USA) were used todetermine total micromolar glycosyl units (glucose, glucose-6-P, andglucose from glycogen) from the clarified samples and from the originalhomogenate (glucose, glucose-6-P only). Results were expressed as mgglycosyl units per g wet tissue.

Although muscle glucose concentrations were not altered by dietarytreatment (FIG. 4, panel A), glycogen and total glycosyl units wereincreased (P=0.05) in piglets of dams fed the a1DHA diet vs. the CONTdiet. In contrast, concentrations were unchanged in piglets of dams fedthe PFO and COCO diets. As with muscle, liver glycogen and totalglycosyl units in piglets from dams fed the a1DHA diet tended to behigher (P=0.089, FIG. 4, panel B). Neither of the (n-3) PUFA diets, northe COCO diet, altered liver glucose concentrations as compared with theCONT.

EXAMPLE 5 RNA Extraction and Quantitative PCR

Total RNA was recovered from cells using Trizol reagent (Invitrogen,Carlsbad, Calif. USA), DNase treated using the Turbo DNase® (Ambion;Houston, Tex. USA) and total RNA (2 μg) was reverse transcribed usingthe iScript cDNA synthesis kit (BioRad; Hercules, Calif. USA).

Primer sequences were: porcine AMPKα2, 5′-cgacgtggagctgtactgat-3′ and5′-cataggtcaggcagaacttgc-3′, porcine SGLT1, 5′-cgtgctgtttccagatgatg-3′and 5′-atcagctccatgaccagctt-3′, and porcine ribosomal protein L32(housekeeper), 5′-tggaagagacgttgtgagcaa-3′ and5′-eggaagtttctggtacacaatgtaa-3′, all sequences are sense and anti-senserespectively.

Thermal cycler conditions for PCR reactions were 95° C. for 3 minfollowed by 40 cycles of 95° C. for 15 s, 65° C. for 30 s, and 72° C.for 30 s. Polymerase chain reaction products amplified were cloned intopGEMT vector (Promega, Madison, Wis. USA) and sequenced forverification. Real-time reactions were carried out on an iCyclerreal-time machine using the IQTM SYBR Green Supermix kit (BioRad,Hercules, Calif. USA). Abundance of gene product was calculated byregressing against the standard curve generated in the same reaction bytheir respective plasmids and gene values normalized to ribosomalprotein L32 (RPL32) housekeeper gene which was not affected by thedietary treatment (P>0.10).

To confirm the increased glucose uptake in (n-3) PUFA treatments and tovalidate a potential mechanism, total GLUT2 and SGLT1 protein expressionand the SGLT1 mRNA abundance in the jejunum was examined. Compared withthe CONT piglet jejunum, semi-quantitative immunoblot analysis showedthat PFO, a1DHA and COCO treatment piglets tended to have greater totalGLUT2 protein expression by approximately 20% (P=0.095, FIG. 2).Moreover, both the PFO and a1DHA diets resulted in significantly highertotal protein expression of SGLT1 as compared with piglets from dams fedthe CONT diet (P<0.05, FIG. 3), whereas there was no effect of the COCOdiet. See also FIGS. 6 and 7. Quantitative PCR was also conducted todetermine whether the mRNA abundance (i.e., log starting quantity) ofSGLT1 was also influenced by maternal diet. Dietary fatty acidsupplementation did not alter SGLT1 mRNA expression in the jejunum (datapresented as log starting quantity for CONT (1.87), PFO (1.71), a1DHA(1.84) and COCO (1.70), pooled SEM-0.14).

EXAMPLE 6 Protein Expression

Whole frozen jejunum sections (1 g) were and homogenized on ice in 700μL Buffer A (50 mmol/L Tris-HCl pH 7.5, 50 mmol/L NaF, 5 mmol/L SodiumPyrophosphate, 1 mmol/L EDTA, 1 mmol/L DTT, 0.1 mmol/LPhenylmethylsulfonyl fluoride, 10% glycerol) containing 1% Triton X-100,5 μmol/L aprotinin, leupeptin, and pepstatin. The lysates werecentrifuged at 6,000×g for 20 minutes at 4° C. to remove insolublematerial. Supernatants were collected and protein was quantified usingBCA reagents (Pierce, Rockford, Ill. USA) and frozen until assayed.Jejunum lysates were used for both the AMPK assay and western blotanalysis.

The abundance of GLUT2 (˜60 kDa) and SGLT1 (˜70 kDa) protein wasdetermined by western blot analysis. Briefly, supernatant containing 250μg protein were immunoprecipitated at room temperature for 2 hours usingthe Catch and Release v2.0 Reversible Immunoprecipitation System(Upstate Cell Signaling Solutions, Charlottesville, Va. USA). Both GLUT2and SGLT1 were immunoprecipitated with 1:100 primary antibody (ChemicomInternational, Temecula, Calif. USA) dilutions. Immunoprecipitatedproteins was separated by SDS-PAGE using a 12% resolving gel andtransferred to a nitrocellulose membrane and probed with primaryantibody for GLUT2 or SGLT1 (1:1000) overnight. The membranes wereprobed with Goat-Anti-rabbit IgG-HRP antibody (Pierce, Rockford, Ill.USA) at 1:20,000 for 1 h at room temperature. Blots were developed usingthe SuperSignal West Pico Chemiluminescent Substrate system (Pierce,Rockford, Ill. USA), captured onto micro-film for analysis anddensitometry of the protein determined using Quantity One 1-D analysissoftware (Bio-Rad, Hercules, Calif. USA).

EXAMPLE 7 Statistical Analysis

All data are presented as means±pooled SEM. The effects of fatty acidswere tested by the PROC MIXED procedure in SAS (Version 9.1, SASInstitute, Cary, N.C.) and treatment differences were evaluated usingleast significant differences, which provided all pair-wise comparisons.Litter/piglet was considered the experimental unit and experimentalreplicate or day of harvest was considered a random effect. Differenceswere deemed significant when P<0.05, and tendencies were noted atP<0.10.

EXAMPLE 8 Animals and Experimental Design

Thirty two females (Ausgene Line 20 dam×SPI sire) were fed one of twoexperimental diets for approximately 150 days to encompass the entiregestation period or 17-19 days over the lactation period before weaning.The dietary treatments (Table 5) consisted of the following: 1) basalcorn/soybean meal control (no added fat, CONT); or 2) the basal dietsupplemented with a protected fish oil product ([PFO], Gromega 365™; JBSUnited Inc., Sheridan, Ind.). The PFO product contains 29% of total fatas n-3 PUFA, with approximately 13% as EPA and 13% as DHA. The gestationand lactation diets were formulated to meet or exceed all therequirements for gestating and lactating sows [NRC, NutritionalRequirements of Swine, 10^(th) ed., Natl. Acad. Press, Washington, D.C.(1998)]. Dams and piglets had access to water at all times.

At farrowing after approximately 150 days on the gestation diets,litters were standardized to ten piglets per dam. To accomplish exposingthe piglets to the n-3 fatty acids only during the gestation or suckling(lactation) period, litters were reciprocally switched such that damsfed the CONT diet received piglets from a darn fed the PFO diet, andvice versa for 15-19 days. The four treatments now consisted ofgestation/lactation feeding to give CONT/CONT, CONT/PFO, PFO/PFO orPFO/CONT piglets. At 15-19 days of age, one medium size piglet (5.4±0.50kg) per litter was randomly transferred to a separate room from thedams, grouped penned and fasted overnight to simulate the weaningprocess (total n=6 per treatment). The following morning, piglets wereeuthanized and tissue samples collected. Small intestinal jejunum andmuscles samples were collected and frozen in liquid nitrogen andadditional jejunum samples placed in 10% formalin for latter analysis.

EXAMPLE 9 Fatty Acid Analysis

Lipids were extracted from piglet muscle and liver by the method ofLepage and Roy [J. Lipid Res., 27: 114-120 (1986), incorporated hereinby reference] with minor modifications. Briefly, 0.5 grams of tissue washomogenized in 2.5 mL 4:1 methanol:hexane and then 200 μL of a 3.7 mmolheptadecanoic acid/L methanol solution added to each sample as aninternal standard. Fatty acid methyl esters were analysed by gaschromatography on a Hewlett-Packard model 6890 (Hewlett-Packard, PaloAlto, Calif.) fitted with a Omegawax 320 (30 m×0.32 mm ID, 0.25 μm)capillary column (Sigma-Aldrich, St Louis, Mo. USA). Hydrogen was thecarrier gas. The temperature program ranged from 80° C. to 250° C. witha temperature rise of 5° C./min. The injector and detector temperatureswere 250° C. and 1 μL of sample was injected and run splitless. Fattyacids were identified by their retention times on the column as judgedfrom appropriate standards.

Piglet Tissues Reflect Dietary Fatty Acid Profiles of Maternal Diet

Fatty acid profiles for the jejunum and longissimus muscle are presentedin Tables 14 and 15. Feeding PFO throughout gestation and lactationresulted in significantly (P<0.05) higher n-3 PUFA contents in both thejejunum and muscle vs. the CONT/CONT regimen. This increase was achievedby both DHA and EPA in the jejunum, but muscle showed enrichment largelyas DHA. Discontinuing the PFO diet at the onset of lactation caused asignificant decrease in the DHA, EPA, and total n-3 contents in bothtissues. However, feeding the PFO diet for the lactation period aloneachieved similar enrichment as did feeding this n-3 source for theentire 150 days.

EXAMPLE 10 Ussing Chamber

Proximal jejunum samples, starting 40 cm from the stomach and consistingof a 20-30 cm segment of the jejunum, were removed and placed in chilledKrebs-Henseleit buffer (pH 7.4), which consisted of the following: 25 mMNaHCO3, 120 mM NaCl, 1 mM MgSO4, 6.3 mM KCl, 2 mM CaCl, 0.32 mM NaH2PO4.The tissue was aerated continuously until clamped in the Ussing chambersin the laboratory. The tunica muscularis was removed from two jejunalsegments per pig, and mounted immediately in Ussing Chambers (DVC 1000World Precision Instruments, New haven, Conn. USA). Each segment wasbathed on its mucosal and serosal surfaces (opening area 1.0 cm2) with 8mL Krebs solution and gassed with 95% O2-5% CO2 mixture. The voltage wasclamped at 0 mV by an external current after correction for solutionresistance. After a 30 minute period to allow the tissues to stabilize,they were challenged with 10 mM D-Glucose added to serosal buffer, andan equimolar concentration of mannitol added to the mucosal buffer.Additionally, jejunum samples from some CONT/CONT piglets were mounted,stabilized and treated (mucosal) with 0.1 mM DHA or 2.5 mM AICARsolubilised in 20 mM taurocholic acid. Glucose uptake was then assessedafter 20 minutes, with the tissues challenged with 10 mM D-glucose asdescribed earlier. The potential difference across the tissue wasmeasured for 30 minutes after each challenge by open circuit conditionsevery 10 seconds due to a short-circuit current being delivered by avoltage clamp apparatus. The change in maximal current was recorded andthe tissue conductance was calculated from the short-circuit current andpotential difference using Ohm's law. This procedure was repeated onfour different days with a pig from each dietary regimen to achieve atotal of four pigs per treatment.

Glucose Transport

Changes in active glucose transport in the jejunum of piglets weanedfrom the dams fed the CONT or PFO diets were compared. As shown in FIG.5, feeding PFO throughout gestation and lactation increased glucoseuptake by 500% (5 vs 25 μA/cm², P<0.05), and providing the n-3 source ingestation alone improved glucose uptake by about 400%. In contrast,feeding PFO in lactation only precluded any significant enhancement inglucose uptake (15 vs 5 μA/cm², respectively, P=0.16).

EXAMPLE 11 Immunoblot Analysis of Glucose Transport Proteins in Totaland Brush Border Membrane Preparations

Fresh intact proximal jejunum was removed, washed with saline and placedon ice while approximately 4 g of mucosa were removed and transferred tocold 2 mM Tris-HCl buffer (pH 7.1) containing 50 mM mannitol andprotease inhibitors (5 μM aprotinin, leupeptin, and pepstatin). Themucosa was then homogenized and PEG 4000 was added to a finalconcentration of 10% and stirred on ice for 15 minutes. The homogenatewas then centrifuged for 15 minutes at 7,500×g and the resultingsupernatant fraction centrifuged at 27,000×g for 60 minutes at 4° C. Thepellet was washed in suspension buffer (10 mM Tris-HCl, pH 7.1,containing 300 mM mannitol and protease inhibitors 5 μM aprotinin,leupeptin, and pepstatin) and collected again by centrifugation for 5minutes, 27,000×g at 4° C. The crude brush border membrane (BBM) pelletwas suspended in 1 mL of suspension buffer. For preparation of totalmembranes, frozen jejunum sections (1 g) were and homogenized on ice in700 μL Buffer A (50 mM Tris-HCl pH 7.5, 50 mM NaF, 5 mM sodiumpyrophosphate, 1 mM EDTA, 1 mM DTT, 0.1 mM phenylmethylsulfonylfluoride, 10% glycerol) containing 1% Triton X-100 and 5 μM aprotinin,leupeptin, and pepstatin. The homogenates were centrifuged at 6,000×gfor 20 minutes at 4° C. to remove insoluble material. The proteinconcentrations of the total and BBM preparations were determined usingBCA reagents (Pierce, Rockford, Ill. USA). Final total and brush bordermembrane preparations were frozen at −80° C. until assayed. The purityof the brush border membrane preparations as measured by alkalinephosphatase were not affected by treatment (data not shown).

The abundance of GLUT2 and SGLT1 protein in total and crude BBM wasdetermined by western blot analysis. Briefly, membrane preparationscontaining 250 μg protein were immunoprecipitated at room temperaturefor 2 h using the Catch and Release v2.0 Reversible ImmunoprecipitationSystem (Upstate Cell Signalling Solutions, Charlottesville, Va., USA).Both GLUT2 and SGLT1 were immunoprecipitated with 1:100 primary antibody(Chemicom International, Temecula, Calif. USA) dilutions.Immunoprecipitated proteins was separated by SDS-PAGE using a 12%resolving gel, transferred to a nitrocellulose membrane, and incubatedwith primary GLUT2 or SGLT1 antibody (1:1000 dilutions) overnight.

Expression of Glucose Transport Proteins in the Jejunum.

Immunoblots for GLUT2 abundance in the crude BBM preparations showed asmall but significant (P<0.05) enrichment in the latter fractionobtained from piglets of dams consuming the PFO diets (FIG. 6, panel A),but there was no apparent change in abundance in total homogenates (FIG.6, panel B). This result was not influenced by duration of the PFOregimen, nor was it specific for the gestation or lactation periods.

Similar immunoblots were performed for SGLT1. As seen in FIG. 7, panelA, the CONT/PFO dietary regimen tended to increased the abundance of theSGLT1 protein in the BBM preparations, but only the PFO/CONT regimen wassignificant (P<0.05). In contrast, feeding PFO in any dietary regimenincreased (P<0.06) the abundance of SGLT1 protein in the totalhomogenate (FIG. 7, panel B).

EXAMPLE 12 Statistical Analyses

All data are expressed as means±SEM. The effects of dietary treatmentregimen were determined by the PROC MIXED procedure is SAS (Version 9.1,SAS Institute, Cary, N.C.) and treatment differences were establishedusing least significant differences procedure when protected by asignificant F-value. The effect of gestation, lactation or gestation andlactation n-3 PUFA feeding was assessed in the model. Litter/piglet wasconsidered the experimental unit and experimental replicate or day ofharvest was considered a random effect. Differences were deemedsignificant at P <0.05, and tendencies are noted at P<0.10.

EXAMPLE 13 Experimental Design

A total of four experimental dietary treatments were employed. Thedietary treatments consisted of 1) basal corn/soybean meal (no addedfat, control), or the basal diet supplemented with either 2) protectedfish oil (Fertilium™ or PFO), 3) a1DHA, and 4) extruded Coconut fat(Coco) (Table 1). The fatty acid profiles of the dietary ingredientsused to provide the fatty acids to the various diets are shown in Table7. The fatty acid composition of all diets used in this experiment werebalanced of the total crude fat percentage of the diets, with the DHApercentage of the DHA diets matched to that of the DHA percentagecalculated in the Fertilium™ diet. Sows were fed a gestation andlactation diet (Tables 1, 2, and 8) continuously starting approximately35-d prior to breeding. Nursery pigs were introduced to starter dietsand taken through a four phase dietary regime (Table 9). Upon exit ofthe nurseries, pigs were phase fed diets (Table 10) containing one ofthe four experimental treatments.

EXAMPLE 14 Animals Sows and Piglets.

Sows were housed in fully enclosed gestation and farrowing rooms whichwere climate controlled. Two hundred forty AusGene genetic sows wereallocated to one of four experimental treatments approximately 35-dprior to breeding. Dry and lactating sows had free access to water atall times and were fed twice daily. Experimental litters were formed bystandardizing litters within treatment. Within treatment, all thepiglets on that farrowing day were pooled and then individually weighedas they were cross-fostered back onto sows of the same treatment.Experimental sow had approximately 10-11 piglets. Individual pigletweights, total number and sex data was recorded for each experimentallitter and the piglets were tattoo and ear notched for treatment, litterand piglet number.

Approximately 2200 piglets were allocated to weaner treatments (Table 9)and weaned to three barns at the production farm the following day.Piglets from sows fed trt 2-4 were evenly split and maintained on eithertheir sow treatment or switch to Coco (if on PUFA diets) or PFO if onCoco diet. Therefore four treatments went to seven consisting of thefollowing: control stayed as the control; PFO, PFO/Coco or PFO/PFO; DHA,DHA/Coco or DHA/DHA and Coco, Coco/PFO or Coco/Coco. Weaned pens aimedto have 24 pigs per pen, equal sex, weight and covariant distributions.Nursery pens were randomly blocked within barns and a total of ninetreatment reps were achieved.

Nursery.

The nursery rooms at the production farm were also thermostaticallycontrolled at the initial temperature of 30° C. and the temperature wasdecreased weekly to a target temperature of 25° C. Piglets had freeaccess to water and were fed ad libitum with a starter pellet diet fortwo weeks before changing to a ground diet. At the end of week 13, pigs,keeping the nursery pens in tacked, were transferred to finisher pennedwithin the same barn.

Grower-Finisher.

Within the finisher barns, pigs had free access to water and were againfeed ad libitum a ground feed containing either control, PFO, DHA orCoco treatment (Table 10).

Overall Growth Performance.

Feed conversion efficiency tended to be improved by up to 2.5% (P<0.10,Table 11) in all PFO or DHA treatments, except for when pigs which hadin utero Coco exposure, compared to the control treatment. Exposure ofpiglets to EPA and DHA in utero and in the sow's milk improved feedconversion in the offspring (P<0.10).

Increase in Piglet Pre-Weaning Growth Rate and Reduced Pre-WeaningMortality when Sows were Fed n-3 PUFA During Gestation and Lactation.

Sows were fed corn/soy diets supplemented with protected n-3 PUFA fromFertilium™ to provide 0.022% DHA and EPA in the final sow feed (Tables12 and 13). Diets were fed to sows for approximately 35-days prior tobreeding and for the entire subsequent gestation and lactation period.Litters were all standardized to the same number of piglets. Number ofpigs and litter weight was collected 14-days post farrowing to determinedietary impact on piglet pre-weaning growth rate and mortality rate. TheFertilium™ diet increased the number of pigs weaned and the pig weaningweight (Tables 12 and 13).

EXAMPLE 15 Statistical Analyses

All data was analyzed by PROC MIXED procedure is SAS (Version 9.1, SASInstitute, Cary, N.C.). In the lactation experiments, sow was consideredthe experimental unit and blocked on week and farrowing room. Fornursery and grower-finisher experimental data, pen was the experimentalunit and was blocked on treatment and by the nine reps generated atweaning.

EXAMPLE 16 Nutrient Transport in Chicks

Nine S1 Leghorn layer hens were housed in individual pens in one sectionof an environmentally controlled facility. Hens were fed diets differingin n-3 polyunsaturated fatty acid (PUFA) and docosahexaenoic acid (DHA)content, and formulated to meet NRC poultry requirements (NationalResearch Council. 1994. Nutrient requirements of poultry. 9^(th) Ed.NRC, Washington D.C.). The three dietary treatments were fed to evaluatethe impact of maternal n-3 PUFA and DHA intake on offspring intestinalnutrient uptake. Treatments included; 1) (CON) Diet supplemented withsoybean oil at a dietary inclusion rate of 2.7%; 2) (PFO) Dietsupplemented with protected fish oil from GROMEGA 365™ (JBS United,Sheridan, Ind.) at 13.56% of the diet to provide a non-algal source ofDHA; or 3) (a1DHA) Diet supplemented with DHA from Schizochytrium algaeat 1.13% of the diet (Table 1). Additionally, the delivery of DHA fromprotected fish oil and algae was formulated to be equal for the PFO anda1DHA treatments (Table 2). Diets were formulated based on an estimateddaily feed intake of 115 g/hen/d.

TABLE 1 Experimental diets^(a) Ingredient, % Control alDHA^(a) PFO^(b)Corn 62.00 65.48 53.63 Soybean meal 24.96 23.03 22.43 Limestone 7.757.75 7.75 Soybean Oil 2.70 0.00 0.00 alDHA^(a) 0.00 1.13 0.00 PFO^(b)0.00 0.00 13.56 Dicalcium phosphate 1.42 1.43 1.46 Vitamin premix 0.500.50 0.50 Salt 0.40 0.40 0.40 DL-Methionine 0.11 0.13 0.12 Mineralpremix 0.10 0.10 0.10 Selenium premix 0.05 0.05 0.05 ^(a)alDHA =Schizochytrium algae ^(b)PFO = protected fish oil

TABLE 2 Formulated eicosapentaenoic acid (EPA) and docosahexaenoic acid(DHA) content of experimental diets and calculated daily intake (g/d)Control alDHA^(a) PFO^(b) Inclusion rate of supplement, % 0.00 1.1313.56 EPA content of supplement, % 0.00 0.99 2.61 DHA content ofsupplement, % 0.00 20.94 1.73 EPA supplemented to hen diet, % 0.00 0.010.35 DHA supplemented to hen diet, % 0.00 0.24 0.24 Estimated dailyintake, g/d 115 115 115 Estimated EPA intake, g/d 0.00 0.01 0.41Estimated DHA intake, g/d 0.00 0.27 0.27 ^(a)alDHA = Schizochytriumalgae ^(b)PFO = protected fish oil

EXAMPLE 17 Animal Protocol: Nutrient Transport in Chicks

Hens were fed the experimental diets for 21 days prior to the collectionof approximately 10 fertile eggs from each of 3 hens/trt for hatching(30 eggs set for hatch per treatment). Post hatch, approximately five,3-day old chicks from each hen were sacrificed for analysis ofintestinal glucose and glutamine uptake (15 chicks/trt). Fertile eggswere incubated at 37.5° C. and 60.4% relative humidity in a commercialegg incubator with automatic egg turning (Jamesway, Model #252), and onday 19 the eggs were transferred to a hatching basket and hatched in theincubator. At hatching, chicks were placed in pre-warmed battery cagesand provided water and a common chick starter diet. At approximately 72hours post-hatch, chicks were euthanized by CO₂ asphyxiation andintestinal jejunum segments were harvested for evaluation of intestinalnutrient absorption.

EXAMPLE 18 Ussing Chamber Protocol: Nutrient Transport in Chicks

Proximal jejunum samples between the bile duct and the yolk-sac wereremoved and placed in chilled Krebs-Henseleit buffer (consisting inmmol/L: 25 NaHCO₃, 120 NaCl, 1 MgSO₄, 6.3 KCl, 2 CaCl, 0.32 NaH₂PO₄; pH7.4) for transport back to the laboratory (<40 min) under constantaeration until clamped in the Ussing chambers. Two jejunal segments perchick were immediately mounted in Ussing Chambers (DVC 4000 WorldPrecision Instruments, New haven, Conn. USA). Each segment was bathed onits mucosal and serosal surfaces (opening area 1.0 cm²) with 3 mL Krebssolution and gassed with 95% O₂-5% CO₂ mixture. The intestinal segmentswere voltage clamped at zero mV by an external current after correctionfor solution resistance. After a short-circuit current was establishedand stabilized (5 to 10 min), basal short-circuit current measurementswere taken using MP100A software (BioPac Systems Inc., Santa Barbara,Calif.). The software allowed real-time measurements of current and thuschanges in current were constantly monitored. After the tissue wasstabilized, they were challenged independently with 10 mmol/L D-Glucoseand 10 mmol/L L-glutamine which was added to serosal buffer, withequimolar (10 mmol/L) mannitol added on the mucosal side. The change inmaximal current was recorded and this was repeated on four differentdays with a total of ten chicks per treatment.

EXAMPLE 19 Fatty Acid Modulation of Nutrient Transport in Chicks

FIG. 8 shows jejunum glucose uptake in three day-old chicks from hensfed a diet enriched with DHA from Schizochytrium algae (a1DHA) orprotected fish oil (PFO). Intestinal glucose transport (10 mM) wasassessed by modified Ussing chamber technique as described above. FIG. 9shows jejunum glutamine uptake in three day-old chicks from hens fed adiet enriched with DHA from Schizochytrium algae (a1DHA) or protectedfish oil (PFO). Intestinal glucose transport (10 mM) was assessed bymodified Ussing chamber technique.

Chicks hatched from hens supplemented n-3 PUFA and DHA from either algal(a1DHA) and non-algal (PFO) sources displayed significantly increasedjejunal glucose uptake compared to chicks hatched from hens notsupplemented with n-3 PUPA or DHA (P<0.05) (FIG. 8). Active glucoseuptake in a1DHA and PFO treatments were 41% and 37% greater than inchicks fed the control diet, respectively. There was no difference inglucose uptake between a1DHA or PFO (P>0.50) (FIG. 8). There was nodifference among treatments for glutamine uptake (P>0.10) (FIG. 9).

EXAMPLE 20 Exemplary Pro Formula

The following are exemplary formulas for a non-algal compositioncomprising omega-3 fatty acids or esters thereof that may be added tothe feed compositions as herein described.

Extruded GroMega Formula #1:

Ingredient % of Product Wheat Flour 65.45 Menhaden Oil 20.00 AlfalfaMeal 8.80 Dry Molases 5.60 Vitamin Pack 0.15

Swine 10-20 lbs/ton complete feed

Ratio DHA:EPA 0.75-1:1

Extruded GroMega Formula #2:

Ingredient % of Product Menhaden Oil 60 Starch Carrier 40 Total 100

-   Swine 5-10 lbs/ton of complete feed-   Ratio DHA:EPA 0.75-1:1

TABLE 1 Sow gestation and lactation diets (as fed basis) GestationLactation CON PFO alDHA Coconut CONT PFO alDHA Coconut Ingredient, %Corn 75.69 75.69 75.69 75.69 64.96 64.9 64.96 64.96 Soybean Meal, 48%18.66 18.66 18.66 18.66 27.74 27.7 27.74 27.74 Premix 4.65¹ 4.65¹ 4.65¹4.65¹ 6.30² 6.30² 6.30² 6.30² Corn Starch 1.00 — 0.86 — 1.00 — 0.86 —Protected fish oil (PFO)³ — 1.00 — — — 1.00 — — Algal DHA (alDHA) — —0.14 — — — 0.14 — Dry coconut fat — — — 1.00 — — — 1.00 Total 100 100100 100 100 100 100 100 Calculated nutrient content, % Crude fat 3.563.78 3.56 4.36 3.45 3.66 3.45 4.25 Crude protein 15.17 15.26 15.17 15.2119.09 19.1 19.09 19.13 Starch 48.86 48.35 48.75 47.87 42.75 42.2 42.6041.77 Metabolizable energy, MJ/kg 13.68 13.68 13.72 13.85 13.56 13.613.51 13.72 Lysine 0.75 0.75 0.75 0.75 1.10 1.10 1.10 1.10 Phosphorus0.77 0.77 0.77 0.77 0.81 0.81 0.81 0.81 Calcium 0.88 0.88 0.88 0.88 0.910.91 0.91 0.91 ¹The premix provided per kg of diet: 662 mg of choline ascholine chloride; 3.35 mg of retinyl acetate; 0.06 mg ofcholecalciferol; 66 mg of vitamin E as alphatocopherol acetate; 1.4 mgof vitamin K as menadione dimethypyrimidinol bisulfate; 0.44 mg ofbiotin; 44 mg of niacin; 24 mg of pantothenic acid; 7 mg of riboflavin;0.03 mg of vitamin B-12; 1.61 mg of folic acid; 0.25 mg of pyridoxine aspyridoxine HCl; 0.48 mg of thiamine; P, 0.43% as monocalcium phosphate;Ca, 0.80% as calcium carbonate; Na, 0.18% as sodium chloride; K, 0.25%as potassium chloride; Mg, 0.02% magnesium Cu, 10 mg as copper sulfate;Fe, 125 mg as iron sulfate; I, 1.26 as potassium iodate; Mn, 60 mg asmanganese sulfate; Se, 0.3 mg as sodium selenite; Zn, 125 mg as zincsulfate. ²Premix provided per kg of diet: 662 mg of choline as cholinechloride; 3.35 mg of retinyl acetate; 0.06 mg of cholecalciferol; 66 ofvitamin E as alphatocopherol acetate; 1.4 mg of vitamin K as menadionedimethypyrimidinol bisulfate; 0.44 mg of biotin; 44 mg niacin; 24 mg ofpantothenic acid; 7 mg of riboflavin; 0.03 mg of vitamin B-12; 1.59 mgof folic acid; 0.25 mg of pyridoxine as pyridoxine HCl; 0.48 mg ofthiamine; P, 0.43% as monocalcium phosphate; Ca, 1.0% as calciumcarbonate; Na, 0.21% as sodium chloride; K, 0.37% as potassium chloride;Mg, 0.06% magnesium; Cu, 10 mg as copper sulfate; Fe, 136 mg as ironsulfate; I, 1.26 as potassium iodate; Mn, 60 mg as manganese sulfate;Se, 0.3 mg as sodium selenite; Zn, 125 mg as zinc sulfate; and 0.08% Lysas lysine HCL. ³Protected fish oil was supplied by JBS United, Inc.

TABLE 2 Sow lactation diet fatty acid composition Diet CONT PFO alDHACOCO (g/100 g total fatty acids) 6:0 0.00 0.00 0.00 0.06 8:0 0.00 0.000.00 1.44 10:0 0.00 0.00 0.00 1.11 12:0 0.00 1.37 1.38 8.97 14:0 0.000.47 0.28 3.82 16:0 15.53 15.56 15.22 14.53 16:1 0.00 0.67 0.00 0.0018:0 3.18 3.20 2.95 3.09 18:1 23.63 21.55 23.19 19.19 18:2(n-6) 54.0752.36 52.74 44.53 18:3(n-3) 3.02 3.05 2.88 2.69 20:0 0.44 0.41 0.44 0.3620:1 0.00 0.27 0.00 0.00 20:3(n-6) 0.00 0.00 0.00 0.00 20:5(n-3) 0.000.58 0.00 0.00 22:5(n-3) 0.00 0.00 0.00 0.00 22:6(n-3) 0.00 0.51 0.800.00 Other 0.12 0.00 0.12 0.20 Saturated 19.27 21.51 21.20 33.58 Total(n-3) 3.02 4.14 3.68 2.69 Total (n-6) 54.07 52.36 52.74 44.53(n-6)/(n-3) 17.90 12.6 14.3 16.5

TABLE 3 Sow milk fatty acid profile following gestation and lactationfeeding of fatty acid modified diets^(1,2) Diet CONT PFO alDHA COCOPooled (g/100 g total fatty acids) SEM 10:0 0.24 0.17 0.18 0.28 0.05912:0 0.27^(a) 0.20^(a) 0.23^(a) 1.30^(b) 0.154 14:0 3.61 2.68 3.14 4.280.586 16:0 33.91 27.59 28.59 34.47 3.091 16:1 10.51 7.84 8.83 10.87 2.5518:0 5.08 5.55 5.49 4.73 0.305 18:1 29.89 39.22 36.69 28.78 4.73318:2(n-6) 13.39 13.08 13.18 12.26 1.640 18:3(n-3) 0.61 0.57 0.56 0.550.053 20:0 0.09 0.06 0.12 0.04 0.061 20:1 0.22 0.42 0.36 0.25 0.144 20:20.25 0.42 0.43 0.30 0.101 20:3(n-6) 0.34 0.07 0.10 0.00 0.203 20:4(n-6)0.50 0.67 0.71 0.82 0.208 20:5(n-3) 0.00 0.07 0.00 0.00 0.040 22:5(n-3)0.00^(a) 0.27^(b) 0.12^(ab) 0.00^(a) 0.069 22:6(n-3) 0.00^(a) 0.24^(b)0.29^(b) 0.00^(a) 0.035 Other 1.07 0.88 0.99 1.06 0.149 Saturated43.31^(bc) 36.25^(a) 37.75^(ab) 45.11^(c) 3.512 Total (n-3) 0.61^(a)1.16^(b) 0.97^(b) 0.55^(a) 0.122 Total (n-6) 14.72 14.41 14.60 13.681.926 (n-6)/(n-3) 24.1^(b) 12.5^(a) 15.4^(a) 24.9^(b) 1.611 ¹Means ofmilk samples collected from 4 sows per treatment. ²Within a row, meanswith superscripts without a common letter differ, P < 0.05.

TABLE 4 Piglet fatty acid composition of longissimus dorsi muscle andproximal jejunum samples taken at weaning^(1,2) Small intestine MuscleCONT PFO alDHA COCO Pooled CONT PFO alDHA COCO Pooled Fatty acid (g/100g total fatty acid) 10:0 0.00 0.00 0.00 0.00 — 0.00 0.00 0.00 0.00 —12:0 0.00 0.00 0.00 0.00 — 0.00 0.03 0.00 0.06 0.043 14:0 0.13^(a)0.28^(ab) 0.36^(b) 0.46^(b) 0.067 1.93 1.78 2.13 2.28 0.355 16:0 20.8420.95 23.15 23.15 1.711 33.51^(c) 29.62^(ab) 28.86^(b) 31.74^(ac) 0.87016:1 1.96 1.43 1.80 2.35 0.297 9.07 6.24 7.81 8.66 1.570 18:0 22.3222.22 23.34 21.64 0.867 10.23 10.25 9.26 10.71 1.270 18:1 13.20 14.1011.63 13.55 2.088 25.02 27.54 31.86 23.54 3.037 18:2(n-6) 21.78 20.7821.60 21.18 1.501 15.69 16.47 14.07 16.58 1.751 18:3(n-3) 0.46 0.13 0.070.21 0.202 0.25 0.21 0.42 0.24 0.152 20:0 0.55 0.65 0.51 0.47 0.193 0.000.00 0.00 0.00 — 20:1 0.00 0.05 0.04 0.00 0.037 0.15 0.25 0.48 0.150.121 20:2 0.30 0.38 0.39 0.27 0.100 0.20 0.37 0.51 0.19 0.144 20:3(n-6)0.67 0.69 0.71 0.66 0.052 0.43 0.34 0.38 0.48 0.130 20:4(n-6) 12.9211.72 11.60 12.25 0.918 2.67 4.48 2.88 4.36 1.121 20:5(n-3) 0.00^(a)0.47^(b) 0.18^(a) 0.10^(a) 0.121 0.00 0.03 0.00 0.00 0.018 22:4 1.591.15 1.06 1.67 0.207 0.49 0.64 0.46 0.81 0.199 22:5(n-3) 0.92^(ab)1.06^(ab) 0.67^(a) 0.78^(ab) 0.124 0.23^(a) 0.82^(b) 0.31^(ab) 0.07^(a)0.178 22:6(n-3) 0.90^(a) 2.54^(b) 2.34^(b) 0.65^(a) 0.338 0.00^(a)0.81^(b) 0.54^(b) 0.00^(a) 0.132 Other 1.47 1.43 0.57 0.63 0.779 0.150.10 0.05 0.14 0.116 Saturated 45.26 45.50 47.84 46.14 45.66^(c)41.73^(ab) 40.25^(a) 44.78^(bc) 1.162 (n-3) 2.28 4.19 3.26 1.74 0.48^(a)1.87^(b) 1.27^(b) 0.31^(a) 0.244 (n-6) 35.41 33.21 33.99 34.17 18.3620.98 16.95 20.94 2.661 (n-6)/(n-3) 15.56 7.93 10.43 19.61 28.11^(a)11.20^(b) 13.40^(b) 28.07^(a) 4.590 ¹Means of four piglets pertreatment. ²Within a tissue and row, means with superscripts without acommon letter differ, P < 0.05.

TABLE 5 Gestation and lactation diets (as fed basis) Gestation LactationControl PFO Control PFO Ingredient, % Corn 75.69 75.69 64.96 64.96Soybean Meal, 48% 18.66 18.66 27.74 27.74 Vitamin/mineral/phytase premix4.65 4.65 6.30 6.30 Corn Starch 1.00 — 1.00 — Protected fish oil (PFO)^(a) — 1.00 — 1.00 Total 100 100 100 100 Calculated nutrient content, %Crude fat 3.56 3.78 3.45 3.66 Crude protein 15.17 15.26 19.09 19.18Lysine 0.75 0.75 1.10 1.10 Phosphorus 0.77 0.77 0.81 0.81 Calcium 0.880.88 0.91 0.91 EPA^(b) — 0.007 — 0.007 DHA^(b) — 0.007 — 0.007 12:0,14:0 and 16:0^(b) — 0.013 — 0.013 Total n-6 fatty acids 1.58 1.58 1.431.43 Total n-3 fatty acids 0.06 0.13 0.07 0.14 n-6:n-3 fatty acid ratio26.70 12.04 20.51 10.11 ^(a) Protected fish oil was supplied by JBSUnited, Inc. ^(b)Calculated percentage of total fat in diet

TABLE 6 Preweaning mortality of litters reared by sows fed protectedfish oil (PFO) in lactation only (control/PFO), gestation only(PFO/Control), or both (PFO/PFO). Treatment (gestation/lactation)*Significance Control/Control^(a) Control/PFO^(b) PFO/PFO^(c)PFO/Control^(d) SEM Gestation Lactation Interaction Litter number 11.310.9 11.0 11.0 0.26 0.97 0.41 0.42 Wean number 9.1 9.3 9.5 9.9 0.490.057 0.76 0.26 Mortality (%) 14.9 9.1 11.0 7.3 2.55 0.14 0.58 0.016*Sow number: superscripts ^(a)= 19, ^(b)= 16, ^(c)= 22 and ^(d)= 21 sows^($)Sum of the born alive and stillborn

TABLE 7 Diet additive ingredient fatty acid analysis profile. Fattyacids presented as a percentage of total fatty acids.* Dietaryingredient* Fatty Acid FERTILIUM ™ alDHA Coconut C10:0 0.258 0.098 6.702C12:0 (lauric) 0.117 0.327 51.754 C14:0 (myristic) 8.662 9.117 19.393C16:0 (palmitic) 18.009 23.127 9.625 C16:1n7 11.190 0.048 0.029 C18:0(stearic) 3.031 0.545 3.010 C18:1n9 3.067 0.077 0.084 C18:1n7 0.1180.132 6.816 C18:2n6 4.382 0.031 2.266 C18:3n6 0.270 0.240 C18:3n3(α-linolenic) 1.476 0.097 0.090 C20:0 0.182 0.167 0.091 C20:1n9 1.4190.046 C20:3n6 0.184 0.409 C20:4n6 (arachidonic) 0.622 2.410 C20:3 0.2420.237 C20:5n3 (EPA) 12.806 1.656 C22:0 0.134 0.075 0.024 C22:4n6 0.1110.085 C22:5n6 0.343 15.736 C22:5n3 (DPA) 2.185 0.405 0.034 C24:0 0.268C22:6n3 (DHA) 12.213 40.940 Total n3 fatty acids 28.680 43.100 0.124Total n6 fatty acids 5.913 18.911 2.266 Total saturated fatty 30.52733.381 87.565 acids n6:n3 ratio 0.21 0.44 21.25

TABLE 8 Experimental diets denoting the calculated balance of addedfatty acids during sow gestation and lactation. Calculations based ofthe total crude fat percentage and DHA in both PUFA diets match to thecontent in the Fertilium diet. Treatment Control FERTILIUM ™ alDHACoconut Gestation diet Crude fat (%) 3.56  3.76 3.62 4.36  EPA, as % offat in diet — 0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.007 —12:0, 14:0, and 16:0 — 0.013 0.006 0.134 as % of Total test fat in diet(%) 0.000 0.266 0.037 0.229 Lactation diet Crude fat (%) 3.45  3.66 3.524.25  EPA, as % of fat in diet — 0.007 0.0005 — DHA, as % of fat in diet— 0.007 0.008 — 12:0, 14:0, and 16:0 — 0.013 0.006 0.144 as % of Totaltest fat in diet (%) 0.000 0.272 0.038 0.236

TABLE 9 The approximate calculated fatty acid balance of the nurseryphase diets. Added fatty acids present were based on the total crude fatpercentage and the DHA content match for both PUFA treatments to that inthe Fertilium. Treatment Control FERTILIUM ™ alDHA Coconut Phase 1 Crudefat (%) 8.41  8.89 8.56 10.76  EPA, as % of fat in diet — 0.007 0.0005 —DHA, as % of fat in diet — 0.007 0.008 — 12:0, 14:0, and 16:0 as % —0.013 0.006 0.139 Total test fat in diet (%) 0.000 0.272 0.039 0.235Phase 2 Crude fat (%) 3.46  3.64 3.52 4.22  EPA, as % of fat in diet —0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.008 — 12:0, 14:0, and16:0 as % — 0.013 0.006 0.139 Total test fat in diet (%) 0.000 0.2720.038 0.236 Phase 3 Crude fat (%) 3.46  3.63 3.52 4.22  EPA, as % of fatin diet — 0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.008 — 12:0,14:0, and 16:0 as % — 0.013 0.006 0.139 Total test fat in diet (%) 0.0000.272 0.038 0.236 Phase 4 Crude fat (%) 3.54  3.71 3.59 4.31  EPA, as %of fat in diet — 0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.008 —12:0, 14:0, and 16:0 as % — 0.013 0.006 0.138 Total test fat in diet (%)0.000 0.274 0.038 0.235

TABLE 10 The approximate calculated fatty acid balance of the grow-Finisher phase diets. Added fatty acids present were based on the totalcrude fat percentage and the DHA content match for both PUFA treatmentsto that in the Fertilium. Treatment Control FERTILIUM ™ alDHA CoconutPhase 5 (nursery exit-45 kg) Crude fat (%) 3.55  3.73 3.61 4.32  EPA, as% of fat in diet — 0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.008— 12:0, 14:0, and 16:0 as % — 0.013 0.006 0.138 of fat Total test fat indiet (%) 0.000 0.272 0.038 0.235 Phase 6 (45-63 kg) Crude fat (%) 3.59 3.77 3.65 4.38  EPA, as % of fat in diet — 0.007 0.0005 — DHA, as % offat in diet — 0.007 0.008 — 12:0, 14:0, and 16:0 as % — 0.013 0.0060.138 of fat Total test fat in diet (%) 0.000 0.274 0.038 0.236 Phase 7(63-82 kg) Crude fat (%) 3.63  3.80 3.68 4.42  EPA, as % of fat in diet— 0.007 0.0005 — DHA, as % of fat in diet — 0.007 0.008 — 12:0, 14:0,and 16:0 as % — 0.013 0.006 0.139 of fat Total test fat in diet (%)0.000 0.275 0.038 0.236 Phase 8 (82-100 kg) Crude fat (%) 3.68  3.863.74 4.48  EPA, as % of fat in diet — 0.007 0.0005 — DHA, as % of fat indiet — 0.007 0.008 — 12:0, 14:0, and 16:0 as % — 0.013 0.006 0.139 offat Total test fat in diet (%) 0.000 0.274 0.038 0.236 Phase 9 (>100 kg)Crude fat (%) 3.73  3.92 3.79 4.55  EPA, as % of fat in diet — 0.0070.0005 — DHA, as % of fat in diet — 0.007 0.008 — 12:0, 14:0, and 16:0as % — 0.013 0.006 0.138 of fat Total test fat in diet (%) 0.000 0.2740.038 0.236

TABLE 11 Cumulative pig performance of gain (ADG), feed intake (ADFI)and feed conversion (FG) for pigs reared by sows fed differentialsources of fatty acids in gestation + lactatation or to the piglet postweaning. Diets were crossed over from the nursery phase. Sow diet ×Nursery/ Dietary Treatment Finisher diet Control × Fertilium × Fertilium× DHA × DHA × Coconut × Coconut × Significance Total Cumulative †Control Coconut Fertilium Coconut DHA Fertilium Coconut SEM Trt Rep ADG(kg/d) 0.50 0.51 0.51 0.52 0.51 0.52 0.52 0.007 0.28 <0.0001 ADFI (kg/d)1.21 1.23 1.22 1.23 1.22 1.27 1.26 0.019 0.27 <0.0001 F:G 2.43 2.39 2.392.37 2.40 2.43 2.44 0.015 0.017 <0.0001 † Denotes the period betweenweaning and market ADG = average daily gain (kg/d), ADFI = average dailyfeed intake (kg/d), F:G = feed conversion ratio (feed to gain)

TABLE 12 The effect of continuous feeding of Fertilium ™ in gestationand lactation on subsequent litter size and piglet body weights. PooledDiet P- Response criteria Control FERTILIUM ™ SEM value¹ Subsequentlitter Sows, n² 336 336 — — Total born, n 11.7 12.1 0.2 0.146 Live born,n 11.1 11.4 0.2 0.197 Birth weight, lbs/pig 3.82 3.81 0.04 0.906Weaning³ Piglets weaned, n 9.5 10.0 0.2 0.066 Weaning weight, lbs/pig12.15 12.53 0.12 0.026 ¹The main effect of diet was evaluated againstthe error term of diet × group interaction. Group refers to a farrowingroom of 28 sows, half per treatment. ²There were a total of 24 groups ofsows that had individual litter information. ³Due to cross-fostering andbump-weaning, piglets weaned (total of 24 groups with all informationavailable) refers to the total number of piglets moved to the nurserydivided by the total number of treatment litters farrowed, and weaningweight (total of 31 groups weaned) refers to the total pounds of pigsmoved to the nursery divided by the total number of pigs moved withineach treatment group.

TABLE 13 The effect of continuous feeding of Fertilium ™ on piglet bodyweights. Response criteria Control FERTILIUM ™ Diet P-value Sows, n 7788 — Standardized litter Litter size, n 11.6 ± 0.2 11.5 ± 0.1 0.843Piglet weight, lbs 3.76 3.76 Covariable d 14 litter Litter size, n 10.4± 0.1 10.7 ± 0.1 0.130 Piglet weight, lbs¹  9.66 ± 0.20 10.24 ± 0.190.05  ¹Including piglet weight at standardization as a covariable (meansadjusted to 3.76 lb for both treatment groups)

TABLE 14 Fatty acid composition of jejunum samples obtained from pigletsweaned from dams fed the control (Cont) and protected fish oil dietaryregimens during gestation and(or) lactation (G/L).^(1,2) Cont/ContCont/PFO PFO/PFO PFO/Cont Fatty acid (g/100 g) 14:0 0.06 0.09 0.05 0.1016:0 19.82 19.31 20.13 21.52 16:1 1.45 1.27 1.24 1.17 18:0 22.14 25.8623.28 20.03 18:1 13.43 12.31 11.64 14.60 18:2n6 21.65 20.28 20.25 20.0518:3n6 0.25 0.16 0.24 0.24 18:3n-3 0.32 0.33 0.34 0.37 20:2 0.42 0.240.22 0.40 20:3n6 0.63 0.46 0.69 0.51 20:4n6 14.66 12.29 13.61 14.7520:5n-3 0.18^(a) 0.73^(b) 0.74^(b) 0.25^(a) 22:4 1.82 1.27 1.17 1.9422:5n-3 1.11^(a) 1.33^(b) 1.38^(b) 1.40^(b) 22:6n-3 0.27^(a) 3.68^(b)4.51^(b) 2.15^(c) Other 1.77 0.39 0.51 0.54 Total 100.00 100.00 100.00100.00 Saturated 42.69 45.64 43.88 42.19 n-3 2.88^(a) 6.06^(c) 6.97^(c)4.17^(b) n6 37.20 33.20 34.79 35.54 n6/n-3 12.91^(a) 5.47^(b) 5.16^(b)8.81^(b) ¹Means of four piglets per treatment. ²Within rows, meanswithout a common letter differ, P < 0.05.

TABLE 15 Fatty acid composition of longissimus muscle samples obtainedfrom piglets of dams weaned from dams fed the control (Cont) andprotected fish oil (PFO) dietary regimens during gestation and(or)lactation (G/L).^(1,2) Cont/Cont Cont/PFO PFO/PFO PFO/Cont Fatty acid(g/100 g fatty acids) 14:0 0.25 0.22 0.16 0.51 16:0 21.41 21.23 20.6420.48 16:1 2.68 2.55 2.44 3.43 18:0 15.77 15.22 14.59 15.82 18:1 13.5112.92 14.94 17.79 18:2n6 26.92 25.45 23.76 23.46 18:3n6 0.00 0.00 0.080.00 18:3n-3 0.39 0.36 0.32 0.39 20:2 0.61 0.59 0.63 0.68 20:3n6 1.051.09 0.93 1.02 20:4n6 13.32 12.20 12.43 12.18 20:5n-3 0.34 0.98 3.290.30 22:4 2.14 1.55 1.48 1.71 22:5n-3 1.52 1.87 1.83 1.44 22:6n-30.00^(a) 1.97^(b) 2.48^(b) 0.70^(c) Other 0.09 0.00 0.00 0.10 Total100.00 100.00 100.00 100.07 Saturated 37.43 38.47 35.39 36.81 n-32.25^(a) 5.18^(bc) 7.92^(b) 2.84^(ac) n6 41.29 38.74 37.20 36.65 n6/n-318.64^(a) 7.55^(b) 5.89^(b) 14.42^(a) ¹Means of four piglets pertreatment. ²Within rows, means without a common letter differ, P < 0.05.

1.-151. (canceled)
 152. A method of increasing the performance of ananimal, the method comprising the steps of administering to the animal afeed composition comprising an algal composition comprising omega-3fatty acids or esters thereof, wherein the omega-3 fatty acids comprisedocosahexaenoic acid and eicosapentaenoic acid, wherein the ratio ofdocosahexaenoic acid to eicosapentaenoic acid in the algal compositionis about 60:1 to about 1:1, wherein the feed composition as a finalmixture comprises about 0.01% to about 60% by weight of the algalcomposition, and wherein the animal is a species of poultry; andincreasing the performance of the animal.
 153. The method of claim 152wherein the feed composition as a final mixture comprises about 0.01% toabout 4.0% by weight of the algal composition.
 154. The method of claim152 wherein the feed composition as a final mixture comprises about0.01% to about 3.0% by weight of the algal composition.
 155. The methodof claim 152 wherein the feed composition as a final mixture comprisesabout 0.01% to about 1.5% by weight of the algal composition.
 156. Themethod of claim 152 wherein the animal is a chicken.
 157. The method ofclaim 152 wherein the feed composition as a final mixture furthercomprises omega-6 fatty acids or esters thereof.
 158. The method ofclaim 152 wherein the feed composition is administered daily to theanimal.
 159. The method of claim 152 wherein the feed composition as afinal mixture further comprises an antioxidant.
 160. The method of claim152 wherein the ratio of docosahexaenoic acid to eicosapentaenoic acidis about 10:1.
 161. The method of claim 152 wherein the ratio ofdocosahexaenoic acid to eicosapentaenoic acid is about 5:1.
 162. Themethod of claim 152 wherein the ratio of docosahexaenoic acid toeicosapentaenoic acid is about 2:1.
 163. The method of claim 152 whereinthe ratio of docosahexaenoic acid to eicosapentaenoic acid is about 1:1.164. The method of claim 152 wherein the animal is a chick.
 165. Themethod of claim 152 wherein the omega-3 fatty acids in the feedcomposition are stabilized by encapsulation.